Preparation of vaccine manures for agricultural soils



Patented Aug. 26, 1952 PREPARATION OF VACCINE MANURIESFOR i AGRICULTURAL SOILS Arthur Becker, Wetzlar, Germany a. Drawing. Application October 17, 1950, Serial No, 190,638. A In Germany October 22, 1949 The present mventionrelates to aprocess for the production of a vaccine manure for agricultural cultivation soils.

Under normal conditions many different bacll) Claims. (Cl. 7l--6) degradation products until a bacterial count of l about 500 milliards per cc, of each medium is attained and are then combined. It is necessary that the added bacteria should conform with the teria contained in animal excrements reach the 5 criteria of the so-called colourful line for inarable soil via stable manure and the like and stance strong gas formation and reduction in neuthere perform important functions on plant tral-red glucose agar, acid formation in litmus growth. Modern intensive methods of artificial milk, definite indcle produc in yp broth treatment of the soil have for various reasons led and motility. The strains of Streptococcus Pyoto an impoverishment of cultivation soils in bac- 1 genes brevis empl y are Obtained y g a terial which are important for life. Owing to the pure culture of these strains, if desired after havprevailing use of artificial fertiliser the soil bac-" ing first subjected it to 'a feeding p y teria were not increased and in many cases even creased addition of decomposed albumens hard injured; The known phenomenon that vegetable ening it by alternate Culture 0 liquid d Solid materials such for example as traw rgots and 15 CUHUTE media of usual composition and by-culture the like, which after prolonged periods in the under unfavourable temperature conditions, manure heap are used as field manure, can be ryi b w n abnormally high (up found almost unchanged in the soil after L z and abnormally low (down to C.) and then culyears, is probably 'due to a shortage of ce11 turing it so long together with diphtheria bacilli," and Cal-boh dmtaaecomposmg mi g i p-roteus bacilli, pneumococci and staphylococci?" and putrefying bacteria. Among the cellulose t l t e foreign Strains e b and carbohydrate-rdecomposiing micro-organisms Whereupen the Culture is Washed Off inlihe 5 4 it is especially the proteus bacteria which are able manner, as for examp t a p y o ical $01115 to cause considerable decomposition of Vegetable tion of common salt. This treatment rendersthe albumens. The existing shortage of bacteria in S eptecoecufi m ogcnels brevis strains e n l y our cultivation soils may; for its part, again be resistant. The added strain of proteus (Bacteri- 1 the cause of the fruits of the soil lacking factors um W' vulgar/e) s pr f bly priv d of its which are important for human and animal abilitytc sw rm y pr l d l r in pheno nourishment or of vitamin-like active substances een'beining mediebeing present only to an insuih'cientamount. It has e found to be advantageous to keep This againbrings' with it various bad effects to approximately the following p p t s for on the health of the animals and moreover leads the miXfiulie 0f t ated bacteria: /3 B. cola, /3 to a reduced separation of strongly vital, repro- SiTeplfOeeceuS iJyOgenes /3 B. fJ S vulductive and functionally strong-bacteriaby the Zaetis e/ iS also Used, p oanimal organism. Thus the vicious. circle is portions for the mixture should be 1:1:1r1. The closed and the impoverishment 13 the m land count is determined in .the Liebreich counting.... in bac teria important for life is accelerated. e 50 h after e e, Or four; culture The object of t present invention 15130 arrest solutions have been combined about 1'70, or 125, this faulty cycle. This is achieved by interruptmimal'ds h bacterium will e contained i ing the cycle at one place and restoring it to a 40 each CC-Qf the mixed culture 501151011- natural and unspoilt cycle between the animal AS examples of r d s, nd th Vegetable 1d h faulty cycle is tion products containing culture solutions I may terrupted by an intensive bacterial treatment of use peptolle Containing b th 01 distilled water the cultivation soil, which is effected by colocontaining p p to ch .5% common salt Risinggr bi la d, garden d, forestry land and 1% dextrose have been added. It is desirable and virgin land, with healthy vital bacteria; to carry out the separate cultures of the three According to the invention therefore I provide different strains of bacteria, which result inlarg-e a process for the production of a vaccina manure scale culture, at as constant a temperature "as" for agricultural cultivation soils in which purepossible, and preferably at an incubator tenrcultured strains of coli bacteria, proteus bacteria perature of about 25-30 C. This mass culture and of Streptococcus pyogenes brevis (Rosenof bacteria takes about 6-8 days. In the case of f" bach) and if desired strains of lactis-aerogenes Bacterium proteus culgare it is continued untilbacteria are separately cultured (for the purpose the bacteria have completely decomposed the al-jl of multiplication) at first'in or on solid and then r -human present in the nutrient solution. '1 v 1 l" in liquid media containing albumen oralbumen- The mixed vaccine obtained bymixingthe three nutrient solutions must be diluted prior to its use for manuring purposes, and this can advantageously be done with physiological common salt solution.

In this case about parts of common salt solution should be used for 1 part of vaccine mixture. The solution can be sprayed as such on to the field, or it .can be added to liquid stable manure before use By treating soils with mixed Vaccines according to the invention and even without additional.

manuring with other substances a far-reaching enrichment of the treated soils with functionally active bacteria is achieved.

For the better understanding of the invention the following example is given tration.

Example One starts with a fairly large number of strains of coli, one strain of non-swarming Baeterz'um. proteusv oulgare and several strains of Streptococcus pyogenes brem's. The two first mentioned are pre-cultured at 37 C. and the streptococcus in Roux dishes at room temperaonly as an illuseach nutrient solution has been reached, the

three fluids, which contain the coli and proteus bacteria, as well as the streptococci, are poured together and grown for a further 24 hours at 371C. The resulting mixed vaccine, in suitably diluted condition is then used for the treatment of soil.

The superiority of a soil manuring with the mixed vaccines according to the invention, over.

an artificial manuring is illustrated in the following table. The table is based on manuring experiments with wheat, barley, and oats, and

for the purpose of comparison, the results obing albumen or albumen degradation products for the purpose of multiplication until a bacterial count of about 500 milliards of bacteria per cc. of nutrient solution is obtained, whereupon the three nutrient solutions are combined and the resulting mixed vaccine is diluted before being applied to the cultivation soils.

2. ,Aprocess as claimed in claim 1 in which the culture on the solid or liquid'nutrient media takes place at a constanttemperature of from i 3. A process as claimed in claim 1 in which the culture of the B. proteus vulgare, which has been artificiallyideprived of its power to swarm, is continued until the albumen contained in the nutrient solution has been completely decomposed.

4. A'process as: claimed in claim 1 in which a peptone broth 'o'ri distilled water containing peptone is utilised as liquid nutrient medium.

5. A process as claimed in claim 1 in which the resulting mixed vaccine is diluted with physiological common salt solution inthe proportion 6. A process as claimed in claim 1 in which strains of B. Lactic 'aerogenes are also used.

8. A process as claimed in claim 6 in which the culture of the B. proteusvulgare, which has been artificially deprived of its power to swarm, is continued until the albumen contained in the nutrient solution has been completely decomposed.

9. A process as claimed in claim 6 in which apeptone broth or distilled water containing peptone'is utilised as liquid nutrient medium.

10. A process as claimed in claim 6 in which the resulting mixed vaccine is diluted with phys- .iological common salt solution in the proportion I AR'I'HUR BECKER.

REFERENCES CITED The following references are of record in the tained without manuring are also included. Number Name Date .Manuringwith- 947,798 'Coates Feb. 1, 1910 Mixed vaccines Artificial Manure Without Manure L.L R. L. R. N. L. L. R.L. R. N. L. L. n. L. R..N.' I

cm, cm. cm. c'm. cm.

941 M 13-19 6-8 as 15-2o.5. 7-9 3-6 an 4-6 l62l.5 7-10 4-6 16-23 6-9 5-6 L'." le ngth' of leaves in cm. R-. -'L. =length of roots in .0111. B; N.=numbcr of roots. V l

Thesowing took place on .30. 4. and the ma-- Number Name Date nuring on-5. 5. The lengths of leaves and roots 1,268,563 Grinberg. June 4,1918 as well as the number of the roots were deter- 1,320,701. Manns Nov..4,.l919. mined-on l6. 5. as the averages of about 50-60 2,043,265 .Roeder June 9, 1936 sample plants. 2,200,532 Bond, May 14, 1940 What I claimis:

l.v A process for the production of an inoculation manure for agricultural soils, in which pure cultured strains of B, coli, B. proteus vulgare and Streptococcus pyogencs brevis (Rosenbach) are separately cultured at first in or on solid and, then in liquid nutrient media contain- OTHER REFERENCES Buchanan, Bacteriology, Fourth Ed; The MacMillanCo, N. Y., pages 397-402.

Baltimore, pages 3374338. 

1. A PROCESS FOR THE PRODUCTION OF AN INOCULATION MANURE FOR AGRICULTURAL SOILS, IN WHICH PURE CULTURED STRAINS OF B. COLI, B. PROTEUS VULGARE AND STREPTOCOCCUS PYOGENES BREVIS (ROSENBACH) ARE SEPARATELY CULTURED AT FIRST IN OR ON SOLID AND THEN IN LIQUID NUTRIENT MEDIA CONTAINING ALBUMEN OR ALBUMEN DEGRADATION PRODUCTS FOR THE PURPOSE OF MULTIPLICATION UNTIL A BACTERIAL COUNT OF ABOUT 500 MILLIARDS OF BACTERIA PER CC. OF NUTRIENT SOLUTION IS OBTAINED, WHEREUPON THE THREE NUTRIENT SOLUTIONS ARE COMBINED AND THE RESULTING MIXED VACCINE IS DILUTED BEFORE BEING APPLIED TO THE CULTIVATION SOILS. 